Composition and method for inhibiting the growth of animal pathogens

ABSTRACT

Isobutyrates in combination with water soluble copper salts are effective in inhibiting the growth of animal and poultry pathogens.

United States Patent [1 1 Field of Search .424/l4l Das [451 Jan. 2, 1973 v [54] COMPOSITION-AND METHOD FOR INHIBITING THE GROWTH OF e ces Cited ANIMAL PATHOGENS OTHER PUBLICATIONS [75] Inventor: Naba columbla Merck Index, eighth edit., (1968) page 302. [73] Assignee: W. R. Grace & Co., New York,

NY. Primary Examiner-Sam Rosen Filed. J 21 1971 Attorney-Kenneth E. Prince 21 Appl. No.: 108,639 [571 ABSTRACT lsobutyrates in combination with water soluble copper [52] U S CI 424/141 424/143. 424/317 7 salts are effective in inhibiting the growth of animal v 51 Int. Cl. .I....A61k 27/00 and poultry paflmgens'. I [58] 9 Claims, No Drawings COMPOSITION AND METHOD FOR INHIBITING THE GROWTH OF ANIMAL PATHOGENS As is well-known in the agricultural industry various animals suffer malfunctioning of the digestive tract with resulting diarrhea. This, of course, leads to poor performance as pertains to weight gain and frequently even death of the animal. A general term applied to this disease is scours. It is uncertain which pathogen is responsible for this condition, and although Escherichia coli is normally found when the condition appears, other pathogens such as Candida, albicans and Sal- A monella species are also involved. It is also well-known that in certain animals diarrhea is a frequent symptom v of Candida albicans infection. Heretofore, then, even though-it has been evident that infections with certain pathogenic organisms were hindering the growth of animals, there has been little success in curing the problem.

As is evident, then, pathogenic organisms which cause malfunctioning of the digestive tract and which ample 9. y I

Examination of Table. 1 reveals that the following when 1.0 percent by weight ammonium isobutyrate is prepared in a liquid medium, the growth of Candida albicans is unaffected. However, when the liquid medium contains 100 pmm CuSO, and 1.0 percent by weight ammonium isobutyrate, the growth of Candida albicans is effectively inhibited as can be seen from Table I, Ex-

ranges of the two components are effective in success fully inhibiting the growth of Candida albicans:

0.5-2.0% by weight-ammonium isobutyrate and 50200,pprn CuSO v I have also found that in addition to the ammonium salt, the potassium, sodium and magnesium salts of "isobutyric acid also are effective in inhibiting the growth of Candida albicans in combination with copper I sulfate or other water-solublecopper salt. Additionally,

, other pathogens whose growthisinhibited by the in-' adversely affect the weight gain of animals cause serious economic losses to the agricultural industry.

Accordingly, it is .an' objective of this invention to provide a means for inhibiting the growth and proliferation of animal pathogens. More specifically, to inhibit the growth of animal pathogens which prevent normal weightgain in animals. It is, more specifically; a furtherobject of this invention to inhibit the growth of Candida albicans, Salmonella species, and Escherichia coli. [t is still a further object to increase the rate of weight gain in animals. Other objectives of this invention will become apparent tothose skilled -in the art from'the material herein disclosed.

l have found that the salts of isobutyric acid in relatively small quantities in combination with water-soluble copper salts inhibits the growth of such aboveponent may still be isobutyric acid and/or the isobutyrate ion. The salts of'isobutyric acid are in general preferred to the acid per se because they have less odor and are more readily stored, shipped and handled. Although the isobutyrate salts in general are effective,

the salts of Groups I and ll of thePeriodic Table as well as theamm'onium salt, are preferred. The sodium and potassium salts are particularly useful and effective.

'The water-soluble copper salts-are generally operable inthis invention. Such salts include, for example, the acetate, bromate, bromide, chlorate, chloride, lactate, nitrate, salicylate, and the like. In general, the cupric salts are more water-soluble than their cuprous counterparts. v I Y r l have found that there is a synergistic effect between isobutyrates and water-soluble copper salts. For example, when 100 ppm CuSO 4 is prepared in a liquid brothmedium and inoculated with Candida albicans, the

growth of Candida albicans is unaffected; Otherwise,

stant invention include Salmonella species and Escherichia coli.

I prefer to treat the infected animaL-particularly poultry, with the compositionof this invention by admixing the poultry feed with 0.5 to 2.0v percent by weight isobutyrate and 50 to ZOO-ppm copper sulfate based on the weight of the composition. When these particular ranges are used, I have found that, in addition to curing this infection, there is also a rate of weight gain increase in the animal being treated as will be seen from the following examples.

The combination described above as added to the site which is to be protected by either dusting, applying a slurr'y,'or spraying of the feedstuffs, 'so that 0.] to 5.0

percent byweight of the total diet will bethe combination heretofore described.

. When administering the pathogen, growth inhibitor to animals I have found several methods of introduction to the animal are satisfactory. In one convenient method the pathogen growth inhibitor is introduced into the animal as a feed supplement by spraying; the pathogen growth inhibitor onto the feedstuff, .so that 0.1-5.0 percent of the animals daily diet contains the pathogen growth inhibitor. Of course, the percentage of pathogen growth inhibitor in the daily'feed will vary with the particular animal. For example, 0.5 to 4.0 percent by weight of the pathogen growth inhibitor in the daily diet is effective for chickens. 1 v

-l have found, therefore, that small quantities of salts of isobutyric acid in combination with water-soluble copper "salts are effective in inhibiting the growth ofpathogenic organisms. Additionally, when animals are fed an additive containing a salt of isobutyric acid and copper sulphate there is a significant weight gain and although .l am uncertain as to why this is so, it is quite possible that the weight gain is a result of the salt in-,

, hibiting the proliferation of pathogens.

7 pathogens in animals is inhibited. The additive come In summation then, l have'found that when 0.1 to 5.0 weight percent (based upon the total weight on an animals daily diet) of an additive is included in an animals daily diet (the weight ratio of additive'to feedstufi is therefore '0.00l-0.05:l), the growth of, certain prises 99.96-99.99 weight percent of an isobutyrate (as defined heretofore herein) and 0.0l-' 0 04 weight percent of a water soluble copper salt (asdefined heretofore). The'weight ratio of the copper salt to isobutyrate which is effective is in the range of 0.000250.0004: 1. Also, 0.5-2.0 percent by weight isobutyrate and 50-200 ppm copper sulfate (based on the weight of the feedstuff) are together efiective to accelerate the growth of the animal eating the feedstuff.

The following examples will aid in explaining the present invention but are intended to be illustrative only, and not limiting.

EXAMPLE 1 (l-a) An agar medium containing l.0percent by weight ammonium isobutyrate and 100 ppm copper sulfate was poured into a petri dish. Candida albicans, a well-known pathogen which was isolated from a chicken exhibiting digestive disorders, was streaked onto the solidified agar. After a 2-week period it was. found that growth of the pathogen had not occurred.

At the same time Candida albicans which was applied to agar medium in petri dishes (l-b) with l ppm copper sulfate but without the ammonium isobutyrate, (l-c) with 1.0% ammonium isobutyrate but without the copper sulfate, and (l-d) without either the ammonium isobutyrate or copper sulfate, each showed substantial growth of pathogen.

EXAMPLES 2-4 Example l-a was repeated except that the potassium (Examplev 2), sodium (Example 3) and magnesium (Example 4) salts of isobutyric acid were substituted for ammonium isobutyrate. Similar results were obtained.

EXAMPLES 5-7 EXAMPLE 8 Example l-a was repeated except that Escherichia coli was substituted forCandida albicans. The results were similar to those attained in Example l-a.

EXAMPLE 9 Liquid broth culture medium containing different combinations of ammonium isobutyrate and copper sulfate were prepared and inoculated with Candida albicans as in Example l-a. Table I below gives the results of the various combinations used and shows the synergistic effect which is attained.

TABLE I In Vitro Inhibitory Effects of Combinations of CuSO4 and Nl'l lsobutyrate on Candida Albicans* CuSO. (ppm).

. NIL-isobutyrate (weight percent) 2$0 100 200 400 800 (The top horizontal row represents susceptibility to CuSO, alone; left vertical column represents susceptibility to NFL-isobutyrate alone. in-

hibitory concentration of mixture: 200 ppm of CuSO. plus 0.5percent of Nl-L-isobutymte. Symbol: growth in tube; no visible growth.

EXAMPLE l0 Triplicate pens of Hubbard white mountain cockerels were reared on a regular commercially available control diet containing 22 percent protein and 2 percent sucrose. The composition of the feed and percentage of each ingredient used were as follows:

Soybean Meal 35.0% Ground Corn 55.63% Salt 0.5% Lime 1.37% Dicalcium phosphate 2.0% Fat v 4.0% Vitamins l.5% Minerals premix (Vitamins A, D;,, D,, E, B Riboflavin, Niacin, D-pantothenic acid, Menadione sodium bisulflte, D -Methionine, Ethoxyquin, Choline chloride.

At the same time, identical triplicate pens were used and dietary additions were made at the expense of the sucrose. One group was therefore, fed a control diet, and a second group was fed substantially the same diet containing 0.1 percent ammonium isobutyrate, a third group was fed substantially the same diet containing 0.5 percent ammonium isobutyrate and 200 ppm copper sulfate, and a fourth group was fed the diet containing 200. ppm copper sulfate, and a fifth group was 'fed the diet containing 2.0 percent ammonium isobutyrate and 50 ppm copper sulfate. The results as seen in Table II show vthat the chickens fed the diets containing both ammonium isobutyrate andcopper sulfate gained more weight than when neither, or only one of, ammonium isobutyrate or copper sulfate was present in the diet.

TABLE II Effects of Ammonium isobutyrate in Grqwing Chickens Dose Response Group No. Treatment Gain Feed g/chick Efficiency 1 Control 728 L55 2 +05% NH4 isobutyrate 696' 1.52 3 +05% NH isobutyrate and 734 L54 200 ppm copper sulfate 4 +200 ppm copper sulfate 727 Lil 5 +20% NH, isobutyrate and 723 1.54

50 ppm copper sulfate Total gain per chick per 4 week experimental Each number is the average of 24 chicks.

"g Feed divided by g gain, average of 3 replications.

What is claimed is:

l. The method of inhibiting the growth of animal pathogens in animals, said pathogens being selected from the group consisting of Candida albicans, Salmon'ella species and Escherichia coli, which comprises adding to the animals daily diet 0.1 to 5.0 weight per cent based upon the total weight of the. animals daily diet of an additive, said additive consisting essentially of 99.96 to 99.99 weight percent of isobutyric acid or a salt thereof and 0.01 to 0.04 weight percent of a watersoluble copper salt.

4; The composition according to claim 3 wherein the isobutyrate is selected from the group consisting of ammonium, calcium, sodium, magnesiu'm,and potassium isobutyrate, and the coppersalt is copper sulfate.

5. A composition of matter according to claim 4 in which the weight ratio of copper sulfate: group member is in the range of 0.00025-0.0004:-l.

6. A composition consisting essentially an animal feedstuff plus an amount of an additive comprising copper sulfate and ammonium isobutyrate effective to accelerate growth of the animal eating the feedstuff.

7. The composition according to claim 6 wherein the feedstuff is selected from the group consisting of ground corn, cracked corn, whole corn, milo, peanut meal, cottonseed meal, wheat, and silage.

8. The composition according to claim 6 wherein the ratio of additive to feedstuff is 0.001 to 0.05:1.

9. The composition according to claim 6 which contains 05 to 2.0 percent by weight isobutyrate and 50 to 200 ppm copper sulfate based upon the'total weight of thecomposition. 

2. The method according to claim 1 in which the isobutyl-rate is selected from the group consisting of ammonium, calcium, sodium, magnesium and potassium isobutyrate and the copper salt is copper sulfate.
 3. The composition comprising a feedstuff and an amount of an additive to inhibit the growth of an animal pathogen selected from the group consisting of Candida albicans, Salmonella species, and Escherichia coli, said additive consisting essentially of 99.96 to 99.99 weight percent of isobutyric acid or a salt thereof and 0.01 to 0.04 weight percent of a water-soluble copper salt.
 4. The composition according to claim 3 wherein the isobutyrate is selected from the group consisting of ammonium, calcium, sodium, magnesium, and potassium isobutyrate, and the copper salt is copper sulfate.
 5. A composition of matter according to claim 4 in which the weight ratio of copper sulfate: group member is in the range of 0.00025- 0.0004:1.
 6. A composition consisting essentially an animal feedstuff plus an amount of an additive comprising copper sulfate and ammonium isobutyrate effective to accelerate growth of the animal eating the feedstuff.
 7. The composition according to claim 6 wherein the feedstuff is selected from the group consisting of ground corn, cracked corn, whole corn, milo, peanut meal, cottonseed meal, wheat, and silage.
 8. The composition according to claim 6 wherein the ratio of additive to feedstuff is 0.001 to 0.05:1.
 9. The composition according to claim 6 which contains 0.5 to 2.0 percent by weight isobutyrate and 50 to 200 ppm copper sulfate based upon the total weight of the composition. 